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Phospha‐Michael Addition as a New Click Reaction for Protein Functionalization
Author(s) -
Lee YanJiun,
Kurra Yadagiri,
Liu Wenshe R.
Publication year - 2016
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201500697
Subject(s) - phosphine , conjugate , chemistry , bioconjugation , acrylamide , click chemistry , surface modification , alkyl , michael reaction , combinatorial chemistry , electrophile , organic chemistry , mathematical analysis , polymer , mathematics , copolymer , catalysis
Abstract A new type of click reaction between an alkyl phosphine and acrylamide was developed and applied for site‐specific protein labeling in vitro and in live cells. Acrylamide is a small electrophilic olefin that readily undergoes phospha‐Michael addition with an alkyl phosphine. Our kinetic study indicated a second‐order rate constant of 0.07 m −1 s −1 for the reaction between tris(2‐carboxyethyl)phosphine and acrylamide at pH 7.4. To demonstrate its application in protein functionalization, we used a dansyl–phosphine conjugate to successfully label proteins that were site‐specifically installed with N ɛ ‐acryloyl‐ l ‐lysine and employed a biotin–phosphine conjugate to selectively probe human proteins that were metabolically labeled with N ‐acryloyl‐galactosamine.